Targeting oncogenes for hepatocellular carcinoma

靶向肝细胞癌的癌基因

基本信息

批准号：
10361475
负责人：
DEVANAND SARKAR
金额：
$ 38.77万
依托单位：
VIRGINIA COMMONWEALTH UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10361475
关键词：
Astrocytes Automobile Driving Binding Biological Assay Cancer Etiology Cell Nucleus Cessation of life Clinical Trials Dendrimers Development Endoplasmic Reticulum FDA approved Genes Hepatocyte Human Human Cell Line Knock-out Knowledge Malignant Epithelial Cell Malignant Neoplasms Membrane Membrane Proteins Messenger RNA Micrococcal Nuclease Molecular Molecular Analysis Monitor Mus N-terminal Oncogenes Oncogenic Pathogenesis Pathway interactions Patients Play Post-Transcriptional Regulation Pre-Clinical Model Preventive Primary carcinoma of the liver cells Proteins Protocols documentation RIPK1 gene RNA RNA Interference Therapy RNA-Induced Silencing Complex Role Scaffolding Protein Signaling Molecule Site Small Interfering RNA System TRAF2 gene Therapeutic Transgenic Organisms Translating Translational Regulation Translations Transmembrane Domain Treatment Efficacy Xenograft procedure advanced disease base combinatorial design effective therapy expectation experimental study in vivo innovation knock-down liver cancer model mRNA Stability mouse model mutant nanocomplexes nanoparticle delivery novel nuclease overexpression patient derived xenograft model pre-clinical protein activation recruit secretory protein targeted treatment therapeutically effective ubiquitin ligase

项目摘要

Summary Hepatocellular carcinoma (HCC), the fifth most common cancer and the second most common cause of cancer-related deaths worldwide, has no effective treatment for advanced disease. The present proposal focuses on two interacting proteins, AEG-1 and SND1, which function as bona fide oncogenes for HCC. AEG-1 and SND1 cooperate to increase RNA-induced silencing complex (RISC) activity where AEG-1 functions as a scaffold protein and SND1 functions as a nuclease. However, there is a gap of knowledge in our understanding of the mechanism(s) driving AEG-1 and SND1 cooperative oncogenic functions. In primary hepatocytes, AEG- 1 is predominantly localized in the nucleus, while in HCC cells AEG-1 is primarily localized in the endoplasmic reticulum (ER) membrane. Preliminary results indicate ER-anchoring is required for AEG-1 oncogenic function. ER-anchored AEG-1 binds specifically to secretory and membrane protein-encoding mRNAs to facilitate their translation. Analysis of SND1 RNA-interactome also identifies membrane protein-encoding mRNAs. Co- localization studies show that both AEG-1 and SND1 are located on ER membrane in HCC cells. Both AEG-1 and SND1 activate NF-κB. ER-anchored AEG-1 functions as a platform for upstream signaling molecules of NF-κB pathway and thus plays an essential role in NF-κB activation. The mechanism by which SND1 activates NF-κB is not known. We hypothesize that in transformed hepatocytes, AEG-1 translocates from the nucleus and anchors into the ER membrane where it recruits SND1 and both cooperate to promote HCC by modulating post-transcriptional regulation of mRNAs in RISC, translational regulation of membrane proteins and activation of NF-κB, AEG-1 and SND1 require each other for optimum functioning and might not exert oncogenic activity alone, and combinatorial inhibition of AEG-1 and SND1 might be an effective therapeutic strategy for HCC. Experiments are designed to interrogate these hypotheses using novel mouse models and targeted nanoplexes delivering siRNA for AEG-1 and SND1. This proposal will contribute to our long-term objectives of identifying key players regulating HCC pathogenesis and translating this knowledge into development of novel and effective targeted therapies. The immediate objective of the proposal is in-depth understanding of the molecular mechanisms by which AEG-1 and SND1 promote HCC and evaluate a combinatorial strategy of inhibiting AEG-1 and SND1 in a mouse model as a potential therapeutic. Thus the proposal has both mechanistic and therapeutic significance and innovation. Successful completion of the proposed studies will establish new targets for developing therapeutics and provide pre-clinical evidence for a targeted protocol.

概括肝细胞癌 (HCC)，第五大常见癌症，也是第二大常见原因世界范围内与癌症相关的死亡，目前尚无针对晚期疾病的有效治疗方法。重点关注两种相互作用的蛋白 AEG-1 和 SND1，它们作为 HCC 的真正癌基因发挥作用。和 SND1 合作增加 RNA 诱导的沉默复合物 (RISC) 活性，其中 AEG-1 充当蛋白质支架和 SND1 作为核酸酶发挥作用然而，我们的理解存在知识空白。驱动 AEG-1 和 SND1 协同致癌功能的机制在原代肝细胞中，AEG- 1 主要定位于细胞核，而在 HCC 细胞中 AEG-1 主要定位于内质初步结果表明 ER 锚定是 AEG-1 致癌功能所必需的。内质网锚定的 AEG-1 与分泌蛋白和膜蛋白编码 mRNA 特异性结合，以促进其发挥作用 SND1 RNA 相互作用组的分析还鉴定了编码膜蛋白的 mRNA。定位研究表明，AEG-1 和 SND1 均位于 HCC 细胞的 ER 膜上。 SND1 激活 ER 锚定的 AEG-1 作为上游信号分子的平台。 NF-κB 通路，因此在 NF-κB 激活中发挥重要作用 SND1 激活的机制。我们未知 NF-κB 在转化的肝细胞中，AEG-1 从细胞核转位。锚定到 ER 膜上，招募 SND1，两者合作通过调节促进 HCC RISC 中 mRNA 的转录后调控、膜蛋白的翻译调控和激活 NF-κB、AEG-1 和 SND1 相互需要才能发挥最佳功能，并且可能不会发挥致癌活性单独抑制 AEG-1 和 SND1 或联合抑制 AEG-1 和 SND1 可能是 HCC 的有效治疗策略。实验旨在使用新颖的小鼠模型和有针对性的方法来质疑这些假设纳米复合物为 AEG-1 和 SND1 提供 siRNA 该提案将有助于我们的长期目标确定调节 HCC 发病机制的关键参与者，并将这些知识转化为新型药物的开发该提案的直接目标是深入了解。 AEG-1 和 SND1 促进 HCC 的分子机制并评估组合策略在小鼠模型中抑制 AEG-1 和 SND1 作为一种潜在的治疗方法。成功完成拟议的研究将具有机制和治疗意义和创新。建立开发治疗方法的新目标，并为目标方案提供临床前证据。